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Calpain-Glo™ Protease Assay Technical Bulletin

Literature # TB344

The Calpain-Glo™ Protease Assay is a homogeneous, luminescent assay that measures calpain I (μ) and II (m) activities. The Calpain-Glo™ Protease Assay provides a succinyl proluminescent calpain substrate, Suc-LLVY-aminoluciferin, in a buffer system optimized for calpain activity and luciferase activity. Adding the single Calpain-Glo™ Reagent to test samples in an “add-mix-measure” format results in calpain cleavage of the substrate and generation of a “glow-type” luminescent signal produced by the luciferase reaction. In this homogeneous, coupled-enzyme format, the signal is proportional to the amount of calpain activity present. The calpain and luciferase enzyme activities reach a steady-state so that the luminescent signal peaks rapidly and achieves maximum sensitivity in 5–15 minutes. The homogeneous Calpain-Glo™ Assay is designed for use with multiwell plate formats, making it ideal for automated high-throughput screening (HTS) of calpain activity and inhibition.

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